U.S. patent application number 10/487066 was filed with the patent office on 2005-06-16 for production and use of a polar lipid-rich fraction containing omega-3 and/or omega-6 highly unsaturated fatty acids from microbes, genetically modified plant seeds and marine organisms.
Invention is credited to Abril, Jesus, Banzhaf, Wulf, Kohn, Gerhard.
Application Number | 20050129739 10/487066 |
Document ID | / |
Family ID | 23117970 |
Filed Date | 2005-06-16 |
United States Patent
Application |
20050129739 |
Kind Code |
A1 |
Kohn, Gerhard ; et
al. |
June 16, 2005 |
Production and use of a polar lipid-rich fraction containing
omega-3 and/or omega-6 highly unsaturated fatty acids from
microbes, genetically modified plant seeds and marine organisms
Abstract
The production and use, and in particular, the extraction,
separation, synthesis and recovery of polar lipid-rich fractions
containing eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA),
docosapentaenoic acid (DPA(n-3) or DPA(n-6)), arachidonic acid
(ARA), and eicosatetraneonoic acid (C20:4n-3) from microorganisms,
genetically modified seeds and marine organisms (including fish and
squid) and their use in human food applications, animal feed,
pharmaceutical applications and cosmetic applications.
Inventors: |
Kohn, Gerhard; (Woerrstadt,
DE) ; Banzhaf, Wulf; (Siefersheim, DE) ;
Abril, Jesus; (Westminster, DE) |
Correspondence
Address: |
SHERIDAN ROSS PC
1560 BROADWAY
SUITE 1200
DENVER
CO
80202
|
Family ID: |
23117970 |
Appl. No.: |
10/487066 |
Filed: |
October 21, 2004 |
PCT Filed: |
May 14, 2002 |
PCT NO: |
PCT/US02/15454 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60290899 |
May 14, 2001 |
|
|
|
Current U.S.
Class: |
424/442 ;
426/601; 514/548 |
Current CPC
Class: |
A61K 31/66 20130101;
A61P 31/04 20180101; A61P 9/00 20180101; A61P 25/00 20180101; C11B
1/10 20130101; A61P 35/00 20180101; A61K 2800/86 20130101; A23D
9/013 20130101; A61P 43/00 20180101; A61P 29/00 20180101; A61P
17/02 20180101; A61P 3/02 20180101; A61P 37/02 20180101; A61P 11/06
20180101; A61P 3/00 20180101; A61P 37/00 20180101; A61P 1/04
20180101 |
Class at
Publication: |
424/442 ;
514/548; 426/601 |
International
Class: |
A23K 001/165; A23K
001/17; A61K 031/225 |
Claims
1. A method for providing a human, animal or aquaculture organism
diet supplement enriched with at least one of eicosapentaenoic acid
(EPA), docosahexaenoic acid (DHA), docosapentaenoic acid
(n-3)(DPAn-3), eicosatetraenoic acid (n-3), docosapentaenoic acid
(n-6)(DPAn-6) or arachidonic acid (ARA) comprising the steps: a)
producing a polar lipid-rich fraction enriched with at least one of
EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA from
genetically modified seeds or marine animals; and b) providing said
polar lipid-rich fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA in a form
consumable or usable by humans or animals.
2. The method of claim 1, wherein the animal is a companion
animal.
3. A method for treating a deficiency in at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA comprising the
steps: a) producing a polar lipid-rich fraction enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA from microbes, genetically modified seeds or marine animals;
and b) providing said polar lipid-rich fraction enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA to treat said deficiency.
4. The method of claim 3, wherein said deficiency results in an
inflammatory condition, an immune system imbalance, a
cardiovascular disease, a developmental deficit related to nervous
system development, a woman's health condition or an infant's
health condition.
5. A method for treating a chronic inflammatory disease state of
the lung comprising the steps: a) producing a purified phospholipid
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA from microbes, genetically
modified seeds or marine animals; b) blending said phospholipid
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA with at least one of EPA-,
GLA- or SDA-rich oils; and c) producing an aerosol comprising the
blend of step (b) for the treatment of said disease states.
6. The method of claim 5, wherein the chronic inflammatory disease
state of the lung is COPD, asthma or cystic fibrosis.
7. A method for the treatment of skin lesions, induced burn,
UV-irradiation or other skin disorders comprising the steps: a)
producing a purified phospholipid fraction enriched with at least
one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA
from microbes, genetically modified seeds or marine animals; b)
blending said phospholipid fraction enriched with at least one of
EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA with at
least one EPA-, GLA- or SDA-rich oil; and c) producing a lotion or
cream for the treatment of said skin disorders.
8. A method for treating cachexia or fat malabsorption comprising
the steps: a) producing a purified phospholipid enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA; b) blending said purified phospholipid enriched with at least
one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA
with at least one other purified phospholipid; c) blending said
purified phospholipid enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA with at least one
DHA, EPA, GLA- or SDA-rich oil; and d) producing a liquid or dried
dietetic product for the treatment of said disease states.
9. The method of claim 8, wherein the cachexia or fat malabsorption
is a result of cancer or Crohn's disease.
10. The method of claim 8, wherein the at least one other purified
phospholipid is obtained from the group consisting of soy, rape
seed, evening primrose, safflower, sunflower, canola, peanut, egg
or mixtures thereof.
11. A method for the treatment of H. pylori-infection of
gastrointestinal tract comprising the steps: a) producing a
purified phospholipid fraction enriched with at least one of EPA,
DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA from
microbes, genetically modified seeds or marine animals; b) blending
said phospholipid fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA with at least one
EPA-, GLA- or SDA-rich oil; and c) producing a fat emulsion or a
dietetic product for the treatment of said disease.
12. A method for providing a fat blend enriched with at least one
of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA
comprising the steps: a) extracting a polar lipid-rich fraction
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid or ARA from microbes, genetically modified
seeds or marine animals; and b) mixing said polar lipid-rich
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA with another oil.
13. The method of claim 12, wherein said another oil is selected
from the group consisting of fish oil, microbial oil, vegetable
oil, GLA-containing oil, SDA-containing oil or and mixtures
thereof.
14. A method for providing a blend of polar lipids enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA comprising the steps: a) extracting a polar lipid-rich fraction
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid or ARA from microbes, genetically modified
seeds or marine animals; and b) mixing said polar lipid-rich
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA with another polar
lipid.
15. The method of claim 14, wherein said another polar lipid is
selected from the group consisting of soy polar lipids, rape seed
polar lipids, sunflower polar lipids, safflower polar lipids,
canola polar lipids, peanut polar lipids or egg yolk polar lipids
and mixtures thereof.
16. A fat blend enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA comprising: a) a polar
lipid-rich fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA from microbes,
genetically modified seeds or marine animals; and b) another
oil.
17. The fat blend of claim 16, wherein said another oil is selected
from the group consisting of fish oil, microbial oil, vegetable
oil, GLA-containing oil, SDA-containing oil and mixtures
thereof.
18. A blend of polar lipids enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA comprising the
steps: a) extracting a EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid or ARA-enriched polar lipid-rich fraction
from microbes, seeds or marine animals; and b) mixing said EPA,
DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA-enriched
polar lipid-rich fraction with another polar lipid.
19. The blend of polar lipids of claim 18, wherein said another
polar lipid is selected from the group consisting of soy polar
lipids, rape seed polar lipids, sunflower polar lipids, safflower
polar lipids, canola polar lipids, peanut polar lipids or egg yolk
polar lipids and mixtures thereof.
20. Purified phospholipids enriched with at least one EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA derived from polar
lipid-rich fraction extracted from genetically modified seeds or
marine animals.
21. The purified phospholipids of claim 20, wherein said EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA-enriched
phospholipid-fraction is in a form consumable or usable by humans
or animals.
22. A method for providing a human, animal or aquaculture organism
diet supplement enriched with at least one of eicosapentaenoic acid
(EPA), docosapentaenoic acid (n-3)(DPAn-3), eicosatetraenoic acid
(n-3) or docosapentaenoic acid (n-6)(DPAn-6) comprising the steps:
a) producing a polar lipid-rich fraction enriched with at least one
of EPA, DPA(n-3), DPA(n-6) or eicosatetraenoic acid from microbes,
genetically modified seeds or marine animals; and b) providing the
polar lipid-rich fraction enriched with at least one of EPA,
DPA(n-3), DPA(n-6) or eicosatetraenoic acid in a form consumable or
usable by humans or animals.
23. A dietetic, pharmaceutical or cosmetic composition comprising a
polar lipid-rich fraction selected from the group consisting of: a)
a polar lipid-rich fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6) eicosatetraenoic acid or ARA from genetically
modified seeds or marine animals, and b ) a polar lipid-rich
fraction enriched with at least one of EPA, DPA(n-3). DPA(n-6) or
eicosatetraenoic acid from microbes genetically modified seeds or
marine animals.
24. A dietetic, pharmaceutical or cosmetic composition comprising
the fat blend of claim 16.
25. A dietetic, pharmaceutical or cosmetic composition comprising
the blend of polar lipids of claim 18.
26. A dietetic, pharmaceutical or cosmetic composition comprising
the purified phospholipids of claim 20.
27. The method of claim 1, wherein said marine animals are fish,
squid, mollusks or shrimp.
28. The method of claim 1, wherein said marine animals are fish or
fish eggs from the group including salmon, tuna, haddock, sardines,
mackerel, or menhaden.
29. The method of claim 3, wherein said microbes are selected from
fungi, microalgae, protozoa or bacteria.
30. The method of claim 3, wherein said microbes are selected from
the group consisting of Stramenopiles, Thraustochytriales,
Chrysophyceae, Xanthophyceae, Bacillariophyceae, Dinophyceae,
Phaeophyceae, Rhodophyceae, Chlorophyceae, Euglenophyceae,
Cryptophyceae, Oomycetes, Chytridomycetes, and Zygomycetes.
31. The method of claim 3, wherein said microbes are from a genus
selected from the group consisting of Mortierella, Mucor,
Phycomyces, Rhizopus, Pythium, Ochromonas, Nitzschia,
Phaeodactylum, Skeletonema, Fucus, Laminaria, Platymonas, Achyla,
Phytophera, Schizochytrium, Thraustochytrium, and
Crypthecodinium.
32. The method claim 1, wherein said at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid, or ARA comprises at
least two weight percent of the total fatty acids of the polar
lipid fraction.
33. The method of claim 1, wherein said at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid, or ARA comprises at
least five weight percent of the total fatty acids of the polar
lipid fraction.
34. The method of claim 3, wherein said seeds or microbes have been
genetically modified to increase their n-3 or n-6 HUFA content.
35. The method of claim 3, wherein said seeds or microbes have been
genetically modified to increase the production of at least one of
EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA.
36. The method of claim 1, wherein said seeds are from a plant
selected from the group consisting of canola, rape seed, linseed,
flaxseed, sunflower, safflower, peanut, soybean and corn.
37. The method of claim 3, wherein said polar lipid-rich fraction
is extracted from said seeds or microbes using alcohol.
38. The method of claim 1, wherein said polar lipid-rich fraction
is derived as a by-product of oil extraction from said seeds using
hexane or other non-polar solvent.
39. The method of claim 3, wherein said polar lipid-rich fraction
is extracted from said seeds or microbes by use of gravity or
centrifugal extraction technology.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to the fields of production
and use, and in particular, the extraction, separation, synthesis
and recovery of polar lipid-rich fractions containing
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA),
docosapentaenoic acid (DPA(n-3) or DPA(n-6)), arachidonic acid
(ARA), and eicosatetraenoic acid (C20:4n-3) from microorganisms,
genetically modified seeds and marine organisms (including fish and
squid) and their use in human food applications, animal feed,
pharmaceutical applications and cosmetic applications.
BACKGROUND OF THE INVENTION
[0002] Highly unsaturated fatty acids of the omega-6 and omega-3
series represent a special class of bioactive lipids in that they
are important structurally in membranes in the body, but also
participate directly and indirectly in communication between cells
through the eicosanoid pathways and by their influence of these
fatty acids on gene expression. Six of these fatty acids,
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA),
docosapentaenoic acid (DPA(n-3) or DPA(n-6)), arachidonic acid
(ARA), and eicosatetraenoic acid (C20:4n-3) have been shown to be
effective in preventing/treating cardiovascular disease,
inflammatory disease, immune function imbalances, fertility and
some of these fatty acids (ARA, DHA DPA(n-6) are important
structurally in the brain and nervous system. Recent evidence
indicates that some highly unsaturated fatty acids may be more
bioavailable when supplied in a phospholipid form than in a
triglyceride form. EPA, DHA, and ARA have historically been
supplied to the nutritional supplement markets in the form of oil
extracted from algae or fish. However recent evidence indicates
that some polyunsaturated fatty acids may be more bioavailable in a
phospholipid form rather than in a triglyceride form. This may be
because of the bipolar nature of phospholipids, making them readily
solubilizable in the gut and available for digestion and uptake.
This same bipolar property of phospholipids additionally would make
these fatty acids more functional in topical applications such as
creams and lotions or more soluble in aqueous-based applications
such as beverages because of there ability to participate in
emulsification processes. We propose that there may be important
advantages in supplying these omega-3 and omega-6 HUFAs in the form
of phospholipids and improved processes for recovering polar lipids
enriched in these fatty acids are also needed.
[0003] Examples of polar lipids include phospholipids (e.g.
phosphatidylcholine, phosphatidylethanolamine,
phosphatidylinositol, phosphatidylserine, phosphatidyl-glycerol,
diphosphatidylglycerols), cephalins, sphingolipids (sphingomyelins
and glycosphingolipids), and glycoglycerolipids. Phospholipids are
composed of the following major structural units: fatty acids,
glycerol, phosphoric acid, and amino alcohols. They are generally
considered to be structural lipids, playing important roles in the
structure of the membranes of plants, microbes and animals. Because
of their chemical structure, polar lipids exhibit a bipolar nature,
exhibiting solubility or partial solubility in both polar and
non-polar solvents. The term polar lipid within the present
description is not limited to natural polar lipids but also
includes chemically modified polar lipids. Although the term oil
has various meanings, as used herein, it will refer to the
triacylglycerol fraction.
[0004] One of the important characteristics of polar lipids, and
especially phospholipids, is that they commonly contain
polyunsaturated fatty acids (PUFAs: fatty acids with 2 or more
unsaturated bonds). In many plant, microbial and animal systems,
they are especially enriched in the highly unsaturated fatty acids
(HUFAs: fatty acids with 4 or more unsaturated bonds) of the
omega-3 and omega-6 series. Although these highly unsaturated fatty
acids are considered unstable in triacylglycerol form, they exhibit
enhanced stability when incorporated in phospholipids.
[0005] The primary sources of commercial PUFA-rich phospholipids
are soybeans and canola seeds. These biomaterials do not contain
any appreciable amounts of highly unsaturated fatty acids unless
they have been genetically modified. The phospholipids (commonly
called lecithins) are routinely recovered from these oilseeds as a
by-product of the vegetable oil extraction process. For example, in
the production of soybean or canola oil, the beans (seeds) are
first heat-treated and then crushed, ground, and/or flaked,
followed by extraction with a non-polar solvent such as hexane.
Hexane removes the triacylglycerol-rich fraction from the seeds
together with a varying amount of polar lipids (lecithins). The
extracted oil is then de-gummed (lecithin removal) either
physically or chemically as a part of the normal oil refining
process and the precipitated lecithins recovered. This process
however has two disadvantages: (1) the seeds must be heat-treated
before extraction with hexane, increasing the processing cost, and
increasing undesirable oxidation reactions and denaturing the
protein fraction, thereby decreasing its value as a by-product; and
(2) the use of the non-polar solvents such as hexane also presents
toxicity and flammability problems that must be dealt with.
[0006] The crude lecithin extracted in the "de-gumming" process can
contain up to about 33% oil (triacylglycerols) along with sterols
and glucosides. One preferred method for separating this oil from
the crude lecithin is by extraction with acetone. The oil
(triacylglycerols) is soluble in acetone and the lecithin is not.
The acetone solution is separated from the precipitate (lecithin)
by centrifugation and the precipitate dried under first a fluidized
bed drier and then a vacuum drying oven to recover the residual
acetone as the product is dried. Drying temperatures of
50-70.degree. C. are commonly used. The resulting dried lecithins
contain approximately 2-4% by weight of oil (inacylglycerols).
Process temperatures above 70.degree. C. can lead to thermal
decomposition of the phospholipids. However, even at temperatures
below 70.degree. C. the presence of acetone leads to the formation
of products that can impair the organoleptic quality of the
phospholipids. These by-products can impart musty odors to the
product and also a pungent aftertaste.
[0007] What is needed is an improved process for effectively
recovering polar lipids and phospholipids rich in omega-3 and
omega-6 HUFAs from biomaterials that enables the use of these fatty
acid in food, nutritional supplement, pharmaceutical and cosmetic
applications. Furthermore the fractions are needed as an ingredient
in feed for companion animals and in aquaculture.
SUMMARY OF THE INVENTION
[0008] In accordance with the present invention, an improved
process is provided for recovering polar lipids enriched in omega-3
and/or omega-6 HUFAs from native biomaterials such as seeds and
microorganisms and the use thereof.
[0009] In one embodiment of the present invention, a method is
provided for providing a human, animal or aquaculture organism diet
supplement enriched with at least one of eicosapentaenoic acid
(EPA), docosahexaenoic acid (DHA), docosapentaenoic acid
(n-3)(DPAn-3), eicosatetraenoic acid (n-3), docosapentaenoic acid
(n-6)(DPAn-6) or arachidonic acid (ARA). The method includes the
steps of producing a polar lipid-rich fraction enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid
and ARA from genetically modified seeds or marine animals; and
providing the polar lipid-rich fraction enriched with at least one
of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA in a
form consumable or usable by humans or animals. Preferably, the
animal is a companion animal.
[0010] In another embodiment of the present invention, a method is
provided for treating a deficiency in at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA. The method
includes the steps of producing a polar lipid-rich fraction
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid and ARA from microbes, genetically modified
seeds or marine animals; and providing the polar lipid-rich
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid and ARA to treat the deficiency.
The deficiency can result in an inflammatory condition, an immune
system imbalance, a cardiovascular disease, a developmental deficit
related to nervous system development, a woman's health condition
or an infant's health condition.
[0011] In another embodiment of the present invention, a method is
provided for treating a chronic inflammatory disease state of the
lung. The method includes the steps of producing a purified
phospholipid fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA from microbes,
genetically modified seeds or marine animals; blending the
phospholipid fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA with at least one
of EPA-, GLA- or SDA-rich oils; and producing an aerosol comprising
the blend for the treatment of the disease states. The chronic
inflammatory disease state of the lung can result in chronic
obstructive pulmonary disease (COPD), asthma or cystic
fibrosis.
[0012] In another embodiment of the present invention, a method is
provided for the treatment of skin lesions, induced bum,
UV-irradiation or other skin disorders. The method includes the
steps of producing a purified phospholipid fraction enriched with
at least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid
and ARA from microbes, genetically modified seeds or marine
animals; blending the phospholipid fraction enriched with at least
one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA
with at least one EPA-, GLA- or SDA-rich oil; and producing a
lotion or cream for the treatment of the skin disorders.
[0013] In another embodiment of the present invention, a method is
provided for treating cachexia and severe fat malabsorption. The
method includes the steps of producing a purified phospholipid
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid and ARA; blending the purified phospholipid
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid and ARA with at least one other purified
phospholipid; blending the purified phospholipid enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid
and ARA with at least one DHA, EPA, GLA- or SDA-rich oil; and
producing a liquid or dried dietetic product for the treatment of
the disease states. The cachexia or severe fat malabsorption can be
a result of cancer or Crohn's disease. Preferably, the at least one
other purified phospholipid is obtained from the group consisting
of soy, rape seed, evening primrose, safflower, sunflower, canola,
peanut, egg and mixtures thereof.
[0014] In another embodiment of the present invention, a method is
provided for the treatment of H. pylori-infection of
gastrointestinal tract. The method includes the steps of producing
a purified phospholipid fraction enriched with at least one of EPA,
DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA from
microbes, genetically modified seeds or marine animals; blending
the phospholipid fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA with at least one
EPA-, GLA- or SDA-rich oil; and producing a fat emulsion or a
dietetic product for the treatment of the disease.
[0015] In another embodiment of the present invention, a method is
provided for providing a fat blend enriched with at least one of
EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA. The
method includes the steps of extracting a polar lipid-rich fraction
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid and ARA from microbes, genetically modified
seeds or marine animals; and mixing the polar lipid-rich fraction
enriched with at least one of EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid and ARA with another oil. Preferably, the
another oil is selected from the group consisting of fish oil,
microbial oil, vegetable oil, GLA-containing oil, SDA-containing
oil or mixtures thereof.
[0016] In another embodiment of the present invention, a method is
provided for providing a blend of polar lipids enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA. The method includes the steps of extracting a polar lipid-rich
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid and ARA from microbes, genetically
modified seeds or marine animals; and mixing the polar lipid-rich
fraction enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid and ARA with another polar lipid.
Preferably, the another polar lipid is selected from the group
consisting of soy polar lipids, rapeseed polar lipids, sunflower
polar lipids, safflower polar lipids, canola polar lipids, peanut
polar lipids or egg yolk polar lipids and mixtures thereof.
[0017] In another embodiment of the present invention, a fat blend
is provided enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA. The fat blend includes a
polar lipid-rich fraction enriched with at least one of EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid and ARA from microbes,
genetically modified seeds or marine animals; and another oil.
Preferably, the another oil is selected from the group consisting
of fish oil, microbial oil, vegetable oil, GLA-containing oil,
SDA-containing oil and mixtures thereof.
[0018] In another embodiment of the present invention, a method is
provided for producing a blend of polar lipids enriched with at
least one of EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA. The method includes the steps of extracting an EPA, DHA,
DPA(n-3), DPA(n-6), eicosatetraenoic acid or ARA-enriched polar
lipid-rich fraction from microbes, seeds or marine animals; and
mixing the EPA, DHA, DPA(n-3), DPA(n-6), eicosatetraenoic acid or
ARA-enriched polar lipid-rich fraction with another polar lipid.
Preferably, the another polar lipid is selected from the group
consisting of soy polar lipids, rapeseed polar lipids, sunflower
polar lipids, safflower polar lipids, canola polar lipids, peanut
polar lipids or egg yolk polar lipids and mixtures thereof.
[0019] In another embodiment of the present invention, purified
phospholipids enriched with at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA derived from polar
lipid-rich fraction extracted from genetically modified seeds or
marine animals are provided. Preferably, the EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA-enriched
phospholipid-fraction is in a form consumable or usable by humans
or animals.
[0020] In another embodiment of the present invention, a method is
provided for providing a human, animal or aquaculture organism diet
supplement enriched with at least one of eicosapentaenoic acid
(EPA), docosapentaenoic acid (n-3)(DPAn-3), eicosatetraenoic acid
(n-3) or docosapentaenoic acid (n-6)(DPAn-6). The method includes
the steps of producing a polar lipid-rich fraction enriched with at
least one of EPA, DPA(n-3), DPA(n-6) and eicosatetraenoic acid from
microbes, genetically modified seeds or marine animals; and
providing the polar lipid-rich fraction enriched with at least one
of EPA, DPA(n-3), DPA(n-6) and eicosatetraenoic acid in a form
consumable or usable by humans or animals.
[0021] The polar lipid-rich fraction of the methods or products of
can be provided as an ingredient of dietetic, pharmaceutical and
cosmetic applications.
[0022] The fat blend of the methods or products of the present
invention can be provided as an ingredient of dietetic,
pharmaceutical and cosmetic applications.
[0023] The blend of polar lipids of the methods or products of the
present invention can be provided as an ingredient of dietetic,
pharmaceutical and cosmetic applications.
[0024] The purified phospholipids of the methods or products of the
present invention can be provided as an ingredient of dietetic,
pharmaceutical and cosmetic applications.
[0025] Preferably, the marine animals of the methods and products
of the present invention are fish, squid, mollusks or shrimp.
Preferably, the marine animals are fish or fish eggs from the group
including salmon, tuna, haddock, sardines, mackerel, or
menhaden.
[0026] Preferably, the microbes of the methods and products of the
present invention are selected from fungi, microalgae, protozoa or
bacteria. More preferably, microbes are selected from the
Stramenopiles, Thraustochytriales, Chrysophyceae, Xanthophyceae,
Bacillariophyceae, Dinophyceae, Phaeophyceae, Rhodophyceae,
Chlorophyceae, Euglenophyceae, Cryptophyceae, Oomycetes,
Chytridomycetes, or Zygomycetes. More preferably, the microbes are
selected from the group of genera consisting of Mortierella, Mucor,
Phycomyces, Rhizopus, Pythium, Ochromonas, Nitzschia,
Phaeodactylum, Skeletonema, Fucus, Laminaria, Platymonas, Achyla,
Phytophera, Schizochytrium, Thraustochytrium, or
Crypthecodinium.
[0027] Preferably, the EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid, ARA or mixture thereof employed in the
methods and products of the present invention makes up at least two
weight percent of the total fatty acids of the polar lipid
fraction.
[0028] Preferably, the EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid, ARA or mixture thereof employed in the
methods and products at the present invention makes up at least
five weight percent of the total fatty acids of the polar lipid
fraction.
[0029] Preferably, the plant seeds or microbes employed in the
methods and products to the present invention have been genetically
modified to increase their n-3 or n-6 HUFA content.
[0030] Preferably, the seeds or microbes used in the methods and
products of the present invention have been genetically modified to
increase the production of at least one of EPA, DHA, DPA(n-3),
DPA(n-6), eicosatetraenoic acid or ARA.
[0031] Preferably, the seeds employed in the methods and products
of the present invention are selected from the group consisting of
canola, rapeseed, linseed, flaxseed, sunflower, safflower, peanuts,
soybeans or corn. Preferably, the polar lipid-rich fraction is
extracted from the seeds and microbes using alcohol.
[0032] In an alternative embodiment of the present invention, the
polar lipid-rich fraction is derived as a by-product (e.g., by
de-gumming) of oil extraction from the seeds using hexane and other
non-polar solvents.
[0033] Preferably, the polar lipid-rich fraction used in the
methods and products of the present invention is extracted from the
seeds and microbes by use of gravity or centrifugal extraction
technology.
DETAILED DESCRIPTION OF THE INVENTION
[0034] Because of their bipolar nature, polar lipids (including
phospholipids) are of significant commercial interest as wetting
and emulsifying agents. These properties may also help make HUFAs
in the phospholipids more bioavailable, in addition to enhancing
their stability. These properties make phospholipids ideal forms of
ingredients for use in nutritional supplements, food, infant
formula, pharmaceutical, and cosmetic applications. Dietary
benefits of phospholipids include both improved absorption and
improved incorporation. Phospholipids also have a broad range of
functionality in the body in that they are important cell membrane
constituents, they are good emulsifiers, they can act as intestinal
surfactants, serve as a choline source and as a source of
HUFAs.
[0035] EPA, DHA, and ARA are normally produced for the nutritional
supplement market and food uses through the extraction (by cooking)
of fish (EPA and DHA) such as menhaden, tuna or salmon, or hexane
extraction of fungal biomass (ARA) from the genus Mortierella. The
phospholipids in both processes are removed in a later degumming
step that produces a waste material comprising a complex mixture of
neutral lipids, sterols, glucosides and phospholipids. This
material is normally sold to the domestic animal feed industry to
dispose of it.
[0036] Besides fish and fungal biomass, there are microbial sources
of DHA and DPA(n-6). For the present invention, useful microbes can
be selected from fungi, microalgae, protozoa or bacteria. These
organisms can be selected from the groups including the
Stramenopiles, Thraustochytriales, Chrysophyceae, Xanthophyceae,
Bacillariophyceae, Dinophyceae, Phaeophyceae, Rhodophyceae,
Chlorophyceae, Euglenophyceae, Cryptophyceae, Oomycetes,
Chytridomycetes, or Zygomycetes. Useful microbes can also be
selected from the group of genera consisting of Mortierella, Mucor,
Phycomyces, Rhizopus, Pythium, Ochromonas, Nitzschia,
Phaeodactylum, Skeletonema, Fucus, Platymonas, Achyla, Phytophera,
Schizochytrium, Thraustochytrium, or Crypthecodinium.
Microorganisms are good sources of phospholipids because they can
be grown in culture in a manner that optimizes phospholipid
production and minimizes triglyceride (oil) production. On the
other hand the methods used in this invention allow both oil and
phospholipids to be recovered separately in forms that can be used
directly in food, feed, nutritional supplements, cosmetic or
pharmaceutical application.
[0037] DHA, EPA and ARA phospholipids can be recovered from fish,
microalgae, or fungi through the degumming process described above.
However as noted this produces a complex material containing many
other compounds including neutral lipids, sterols, glucosides,
etc
[0038] A preferred embodiment of the present invention is to use
alcohol and centrifugation to recover the omega-3 and/or omega-6
HUPA-rich phospholipids. Preferred methods for this recovery are
described in the following references, which are incorporated by
reference herein in their entirety:
[0039] i. PCT Application Serial No. PCT/US01/12047, entitled
"Method for the Fractionation of Oil and Polar Lipid-Containing
Native Raw Materials" filed Apr. 12, 2001;
[0040] ii. PCT Application Serial No. PCT/US01/12049, entitled
"Method For The Fractionation Of Oil And Polar Lipid-Containing
Native Raw Materials Using Water-Soluble Organic Solvent And
Centrifugation" filed Apr. 12, 2001.
[0041] Once the omega-3 and/omega-6 rich phospholipid fractions
have been extracted by these preferred processes, they can be used
directly as ingredients or they can be purified further and even
separated into phospholipid classes by well-known techniques such
as different forms of chromatography, molecular distillation, and
special refining techniques. Polar lipid groups
phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine,
phosphatidylinositol, and sphingolipids. The phospholipid rich
polar lipids or the purified phospholipid rich fractions can also
be mixed with another lipid or oil such as fish lipids, microbial
lipids, vegetable lipids, GLA-containing lipids, SDA-containing
lipids and mixtures thereof, or be mixed with another phospholipid
fraction (lecithin) such as soy or egg yolk lecithin, sunflower
lecithin, peanut lecithin or mixtures thereof prior to use as a
nutritional supplement, feed or food ingredient. These mixtures of
phospholipids can also be incorporated into creams or lotions for
topical applications (e.g. treating of skin conditions) or skin
lesions induced by burns, UV-irradiation or other skin damaging
processes. The mixtures can also be processed to produce a liquid
or spray-dried dietetic product or fat emulsion for treating
cachexia and severe fat malabsorption or for treatment of H. pylori
infection of the gastrointestinal tract or incorporated into
aerosol sprays for treating chronic inflammatory disease states of
the lung (e.g., COPD, asthma, cystic fibrosis).
[0042] Advantages of the present invention including providing
omega-3 and/or omega-6 HUFAs in a more bioactive and functional
form (phospholipid) than the triglyceride form and include a better
process (e.g., a) no need for heat treatment; b) no use of toxic
solvents (like hexane) and c) no artifacts and off-flavors due to
the use of acetone) for recovering these phospholipids from
oilseeds and microbes. The EPA, DHA, DPA(n-3), DPA(n-6),
eicosatetraenoic acid and/or ARA content of the total fatty acids
of the polar lipid fractions and comprise at least two weight
percent of the total fatty acids of the polar lipid fraction, more
preferably greater than 5 weight percent, more preferably greater
than 10 weight percent, and most preferably greater than 20 weight
percent of total fatty acids in the polar lipid fraction.
EXAMPLE
Example 1
[0043] Phospholipids were extracted from salmon, salmon roe, Black
tiger prawns, squid, Schizochytrium sp, and the total fatty acid
content of the phospholipids was determined by gas chromatography.
The results are presented in Table 1. As can be observed the
phospholipid fraction of these biomaterials can be used to deliver
omega-3 and/or omega-6 HUFAs and in this form these bioactive fatty
acids should be more stable, more bioavailable, and more
functional.
1TABLE 1 Total fatty acid content of phospholipids extracted from 4
types marine animal products and forma microbial source. Salmon
Tiger Salmon roe Prawn Squid Schizochytrium PL's PL's PL's PL's
PL's COMPOUND % TFA % TFA % TFA % TFA % TFA MYRISTATE C14:0 2.99
4.58 1.14 1.87 2.66 MYRISTOLEATE C14:1 0.00 0.00 0.05 0.03 0.00
PALMITATE C16:0 26.48 29.661 25.44 30.37 28.25 PALMITOLEATE C16:1
2.86 4.38 1.55 0.23 0.62 STEARATE C18:0 5.74 6.19 13.59 4.41 1.75
OLEATE C18:1 9.74 13.36 13.33 1.86 6.99 LINOLEATE C18:2n6 3.14 1.05
12.51 0.32 2.46 GAMMA LINOLENATE C18:3n6 0.00 0.00 0.00 0.00 1.09
ARACHIDATE C20:0 0.00 0.00 0.74 0.13 0.00 LINOLENATE C18:3n3 0.49
0.22 0.54 0.05 0.00 OCTADECATETRAENOATE C18:4 0.50 0.33 0.08 0.07
0.00 EICOSENOATE-11 C20:1 0.75 1.07 0.85 5.54 0.00
EICOSADIENOATE-11,14 C20:2 0.00 0.00 0.56 0.34 0.00 BEHENATE C22:0
0.00 0.00 1.35 0.08 0.00 EICOSATRIENOATE C20:3n3 1.01 3.58 0.07
0.42 1.61 ARACHIDONATE C20:4n6 0.89 0.56 5.54 1.70 1.18 ERUCATE
C22:1 0.21 0.00 0.04 1.41 0.00 EICOSAPENTAENOATE C20:5n3 9.50 9.39
8.47 14.71 5.11 LIGNOCERATE C24:0 0.00 0.00 0.56 0.00 0.00
NERVONATE C24:1 1.61 1.65 0.73 0.82 0.00 DOCOSAPENTAENO- C22:5n6
0.00 0.00 0.45 0.22 17.68 ATE n-6 DOCOSAPENTAENO- C22:5n3 3.16 4.40
0.55 0.44 0.00 ATE n-3 DOCOSAHEXAENOATE C22:6n3 30.92 19.50 11.86
35.54 30.61 100.00 100.00 100.00 100.00 100.00
[0044] The present invention, in various embodiments, includes
components, methods, processes, systems and/or apparatus
substantially as depicted and described herein, including various
embodiments, subcombinations, and subsets thereof. Those of skill
in the art will understand how to make and use the present
invention after understanding the present disclosure. The present
invention, in various embodiments, includes providing devices and
processes in the absence of items not depicted and/or described
herein or in various embodiments hereof, including in the absence
of such items as may have been used in previous devices or
processes, e.g., for improving performance, achieving ease and/or
reducing cost of implementation.
[0045] The foregoing discussion of the invention has been presented
for purposes of illustration and description. The foregoing is not
intended to limit the invention to the form or forms disclosed
herein. Although the description of the invention has included
description of one or more embodiments and certain variations and
modifications, other variations and modifications are within the
scope of the invention, e.g., as may be within the skill and
knowledge of those in the art, after understanding the present
disclosure. It is intended to obtain rights which include
alternative embodiments to the extent permitted, including
alternate, interchangeable and/or equivalent structures, functions,
ranges or steps to those Claimed, whether or not such alternate,
interchangeable and/or equivalent structures, functions, ranges or
steps are disclosed herein, and without intending to publicly
dedicate any patentable subject matter.
* * * * *